RESUMO
Acute food withdrawal reversibly inhibits the hypothalamic-pituitary-gonadal axis in men and in rhesus monkeys, and it produces defects in LH pulsatility in normal-weight women. However, the clinical effect of short-term nutritional deprivation on the reproductive axis of normally cycling women has not been evaluated. Thus we studied the effect of a 3-day fast during the midfollicular phase on menstrual cycle length, gonadotropin secretory patterns, follicular development, and ovulation. After a baseline ovulatory cycle, 12 women within 15% of ideal body weight were randomized to be fed (n = 7) or fasted (n = 10) on cycle days 7 to 9. Five of the women repeated the study and received the alternate diet. Endocrine and metabolic parameters of fasting and reproductive physiology were measured on cycle days 6 to 12. Fasted physiology was demonstrated by characteristic alterations in growth hormone, insulin-like growth factor I, TSH, and T3 levels. During fed cycles, the number of LH pulses remained constant on cycle days 6, 9 and 11, whereas mean LH levels, LH area under the curve, and LH pulse amplitude increased significantly over this time (all P < 0.05). In contrast, fasted cycles were marked by a significant decrease in the number of LH pulses on the last day of the fast (cycle day 9, P < 0.05) and by a lack of increase over time of mean LH values, LH area under the curve, and LH pulse amplitude. Follicle development, as assessed by daily ultrasound examination and estradiol measurements, was similar in all cycles and was followed by ovulation in all women; follicular and luteal phase lengths of fasted and fed cycles were similar. We conclude that the alterations in LH secretory dynamics that occur during a 3-day fast are not sufficient to perturb follicle development and cycle lengths in normal-weight sedentary women. The resilience of the reproductive axis in these healthy women contrasts with the sensitivity of the hypothalamic-pituitary-gonadal axis to acute nutritional withdrawal in men and in monkeys. We speculate that differences in the set point for sensing fuel availability or in the status of energy balance before the initiation of food deprivation may account for these species-specific and sex-specific variabilities.
Assuntos
Peso Corporal , Jejum , Hormônio Luteinizante/metabolismo , Reprodução/fisiologia , Adulto , Feminino , Fase Folicular , Humanos , Sistema Hipotálamo-Hipofisário/fisiologia , Ovário/fisiologia , Valores de Referência , Fatores de TempoRESUMO
The postpartum period is characterized hormonally by elevated levels of PRL and low levels of gonadotropins and sex steroids. In breast feeding, this state of postpartum amenorrhea can persist for an extended period, even though PRL levels decrease slowly. Although the action of PRL on multiple target sites has frequently been suggested as the cause of this ovarian quiescence, a suckling-induced alteration in hypothalamic gonadotropin-releasing hormone (GnRH) production has also been hypothesized. To test this latter hypothesis, we provided a uniform pulsatile GnRH stimulus to eight exclusively breast-feeding women for an 8-week duration beginning at 4 weeks postpartum. Five women with functional hypothalamic amenorrhea served as a comparison group. All women received GnRH administered at a dose of 200 ng/kg every 90 min sc via a portable infusion pump. Serial blood sampling for LH, FSH, and PRL was performed weekly for 5 h at 10-min intervals beginning immediately before initiation of GnRH, during the period of GnRH, and 1 week after the cessation of GnRH. The women collected daily urine aliquots for estrone-3-glucuronide, pregnanediol-3-glucuronide, and LH determinations. Serial transvaginal sonography was used to monitor follicular development. Before GnRH treatment the urinary steroid and serum gonadotropin levels of the two groups were low and similar. As expected, PRL levels were higher in the postpartum women (87 micrograms/mL vs. 4.25 micrograms/L, P < 0.05). After initiation of pulsatile GnRH, LH values increased and FSH values decreased in both groups. The LH increase with GnRH was significantly greater in the breast-feeding group than in the hypothalamic amenorrhea group (19.75 mIU/mL vs. 12.34 mIU/mL, P < 0.05). Analysis of pulse frequency and amplitude revealed a nearly complete 1:1 induction of LH pulses by the exogenous GnRH in both groups, with the breast-feeding group showing a greater amplitude (12.26 mIU/mL vs. 5.34 mIU/mL, P < 0.05). The cycle lengths, urinary steroids, and vaginal ultrasonography demonstrated a more rapid initial ovarian responsiveness in the breast-feeding group, as determined by the length of the first follicular phase. The breast-feeding group also showed a brisker ovarian response, as evidenced by a greater number of follicles that were 12 mm or greater (2.3 vs. 1.2, P < 0.05), and a greater luteal phase peak and integrated pregnanediol excretion, respectively (3.02 micrograms/L creatinine and 39.87 micrograms/L creatinine/cycle vs. 1.89 micrograms/L creatinine and 7.69 micrograms/L creatinine/cycle, P < 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Amenorreia/fisiopatologia , Aleitamento Materno , Hormônio Liberador de Gonadotropina/uso terapêutico , Lactação/fisiologia , Hormônio Luteinizante/metabolismo , Ciclo Menstrual/efeitos dos fármacos , Ovário/efeitos dos fármacos , Período Pós-Parto/fisiologia , Adulto , Estradiol/sangue , Estrona/análogos & derivados , Estrona/urina , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Foliculoestimulante/metabolismo , Humanos , Recém-Nascido , Hormônio Luteinizante/sangue , Ovário/diagnóstico por imagem , Ovário/fisiopatologia , Gravidez , Pregnanodiol/análogos & derivados , Pregnanodiol/urina , Progesterona/sangue , Prolactina/sangue , Valores de Referência , UltrassonografiaRESUMO
Serum concentrations of progesterone begin to rise just before the midcycle gonadotropin surge that leads to ovulation. To examine the role of progesterone in the regulation of these events, we evaluated the effects of a low dose (1 mg/day, orally) of the antiprogesterone RU 486 on the timing of the gonadotropin surge and ovulation in normally cycling women. The drug or a placebo was given for 5 or 15 days, starting when the dominant follicle reached 14-16 mm. RU 486 consistently delayed the timing of the midcycle gonadotropin surge and the subsequent collapse of the dominant follicle, despite rising estradiol concentrations and normal follicular development. Unexpectedly, RU 486 also delayed the emergence of the periovulatory progesterone rise. The addition of progesterone (5-10 mg/day, im, for 2 days) to a 5-day course of RU 486 after the emergence of a mature follicle readily induced LH and FSH surges and completely reversed the effects of RU 486 at midcycle. Our results suggest that RU 486 delays the midcycle gonadotropin surge and ovulation by suppressing or antagonizing an ovarian progestational signal. Progesterone may, thus, represent the ultimate ovarian signal to the estrogen-primed hypothalamic-pituitary unit to trigger the gonadotropin surge that leads to ovulation.
Assuntos
Ciclo Menstrual/efeitos dos fármacos , Mifepristona/farmacologia , Progesterona/metabolismo , Adulto , Estradiol/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Fase Folicular/efeitos dos fármacos , Humanos , Fase Luteal/efeitos dos fármacos , Hormônio Luteinizante/sangue , Ciclo Menstrual/fisiologia , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/fisiologia , Progesterona/sangue , Progesterona/farmacologia , Valores de ReferênciaRESUMO
OBJECTIVE: To determine whether serum levels of placental protein 14, a major product of the progesterone-induced secretory endometrium, accurately reflect histologic maturation of the endometrium. METHODS: Daily serum levels of placental protein 14 were compared in 50 normally cycling women with normal or delayed endometrial maturation, as assessed by histologic dating of an endometrial biopsy in the midluteal phase of the same cycle. Ten of these subjects had placental protein 14 measurements but no biopsy in an additional cycle to examine the potential effects of the biopsy on secretion of this protein. RESULTS: Serum placental protein 14 concentrations started to increase 8 days after the LH surge and peaked at similar levels on the first day of the next menses in biopsy and non-biopsy cycles. The biopsy cycles had a shorter luteal phase but a slightly faster increase in placental protein 14 concentrations. Both the integrated secretion of this protein and single measurements on the day of the biopsy or at the onset of the next menses overlapped substantially in women with different degrees of endometrial development, even when differentiation of the endometrium was severely delayed. CONCLUSION: Serum measurements of placental protein 14 do not accurately predict, and thus should not replace, histologic evaluation of the endometrium at nidation.
Assuntos
Endométrio/citologia , Glicoproteínas , Fase Luteal/fisiologia , Proteínas da Gravidez/sangue , Aborto Habitual/diagnóstico , Adulto , Biópsia , Endométrio/fisiologia , Feminino , Glicodelina , Humanos , Infertilidade Feminina/diagnóstico , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Gravidez , RadioimunoensaioRESUMO
OBJECTIVE: To investigate whether the antiprogestin RU486 acts primarily on the hypothalamus to delay the midcycle gonadotropin surge and thus gain insight into the site(s) of action of P in the control of ovulation. DESIGN: Prospective, crossover, single-blinded clinical study. SETTING: Outpatient clinic in an academic research environment. PATIENTS: Women with hypothalamic amenorrhea. INTERVENTIONS: RU486 or a placebo was given orally at a low dose of 1 mg/d for 5 days, starting when the dominant follicle reached 14 to 16 mm, to women with hypothalamic amenorrhea undergoing ovulation induction with GnRH pulses of unvarying frequency and dose. Blood samples and ovarian ultrasounds were obtained daily in the late follicular phase and every 3 to 4 days in the remainder of the cycle. MAIN OUTCOME MEASURES: Follicular diameter and plasma levels of LH, FSH, E2, and P. RESULTS: RU486 consistently delayed the timing of the midcycle gonadotropin surge and ovulation. Gonadotropin and steroid levels were suppressed during RU486 treatment, but follicular growth progressed normally in most patients. CONCLUSIONS: RU486 does not act primarily on the hypothalamus to delay ovulation. Rather, this compound appears to antagonize P at the pituitary level to suppress gonadotropin and steroid hormone secretion. P may thus act on the pituitary, independent of any hypothalamic effects, to regulate the timing of the midcycle gonadotropin surge and ovulation.
Assuntos
Hormônio Liberador de Gonadotropina/farmacologia , Ciclo Menstrual/efeitos dos fármacos , Mifepristona/farmacologia , Ovulação/efeitos dos fármacos , Progestinas/antagonistas & inibidores , Adulto , Amenorreia/induzido quimicamente , Amenorreia/fisiopatologia , Animais , Feminino , Hormônios Esteroides Gonadais/metabolismo , Humanos , Hipotálamo/efeitos dos fármacos , Folículo Ovariano/efeitos dos fármacos , Placebos , Fatores de TempoRESUMO
OBJECTIVE: To investigate whether luteal and endometrial abnormalities occur more frequently in an infertile population and thus contribute to infertility. DESIGN: Prospective controlled clinical study. SETTING: Outpatient clinic in an academic research institution. PARTICIPANTS: Thirty-three fertile controls and 31 infertile women without ovulatory disorders, tubal disease, or male factors. INTERVENTIONS: All women underwent an endometrial biopsy 9 days after the LH surge followed by an IM injection of 5,000 IU hCG. Blood samples were drawn immediately before hCG administration for serum P and placental protein 14 (PP14) measurements, at 6 hours after hCG stimulation for serum P concentrations, and on day 5 after hCG administration for serum PP14 levels. MAIN OUTCOME MEASURES: Histologic dating of the endometrium and serum P and PP14 measurements. RESULTS: Abnormal endometrial biopsies occurred more frequently in infertile (43%) than in fertile women (9%). Except for one case, these specimens were not associated with low hCG-stimulated P levels. Serum PP14 measurements varied widely and did not discriminate subjects with abnormal endometrial development. CONCLUSIONS: Disruption of endometrial maturation without a concomitant defect of the corpus luteum occurs more frequently in an infertile population and thus may contribute to infertility.
Assuntos
Corpo Lúteo/patologia , Endométrio/patologia , Glicoproteínas/sangue , Infertilidade Feminina/sangue , Infertilidade Feminina/patologia , Proteínas da Gravidez/sangue , Progesterona/sangue , Adulto , Biópsia , Gonadotropina Coriônica , Endométrio/fisiopatologia , Feminino , Glicodelina , Humanos , Infertilidade Feminina/fisiopatologia , Ciclo Menstrual , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de TempoRESUMO
OBJECTIVE: To examine whether midluteal phase administration of the luteotrophic hormone hCG can result in higher and more stable serum levels than random sampling of P and placental protein 14 (PP14). DESIGN: Prospective controlled clinical study. SETTING: Normal human volunteers in an academic research environment. PARTICIPANTS: Twenty-six fertile, regularly cycling women. INTERVENTIONS: Blood samples were drawn at 0, 3, 6, 9, 12, 18, and 24 hours and then daily for the next 6 days, after a single IM injection of 5,000 IU hCG or saline given on day 5, 7, or 9 after the LH surge, as detected by rapid plasma assays. MAIN OUTCOME MEASURES: Serum P and PP14 measurements. RESULTS: Peak P and PP14 concentrations occurred at 6 hours and 5 days, respectively, after hCG stimulation on luteal phase day 9. Progesterone but not PP14 levels were significantly higher and less variable after hCG than after saline administration on this day. Progesterone responses exceeded 11.0 ng/mL (35.0 nmol/L) in all women, suggesting that this represents the cutoff limit for normal luteal function. Because PP14 responses were highly variable and inconsistent, it was not possible to determine a threshold for normal endometrial function. CONCLUSIONS: Midluteal phase administration of hCG in normal women induces consistent serum P levels > 11.0 ng/mL (35.0 nmol/L) but highly variable PP14 responses.
Assuntos
Gonadotropina Coriônica/farmacologia , Glicoproteínas , Fase Luteal/efeitos dos fármacos , Proteínas da Gravidez/sangue , Progesterona/sangue , Adulto , Gonadotropina Coriônica/administração & dosagem , Feminino , Glicodelina , Humanos , Cinética , Estudos ProspectivosRESUMO
OBJECTIVE: To investigate changes in menstrual cycle hormones and endometrial maturation that may contribute to the decline in fertility with aging. DESIGN: Prospective controlled clinical study. SETTING: Normal human volunteers in an academic research institution. SUBJECTS: Women with regular menstrual cycles. INTERVENTIONS: Thirty-two women, aged 20 to 30 or 40 to 50 years, had daily blood drawing starting on cycle day 6 to 10 and continuing until 2 days after the onset of next menses. In addition, 60 women, aged 20 to 30 or 40 to 50 years, had a total of 93 endometrial biopsies performed on day 7 to 9 after the LH surge. MAIN OUTCOME MEASURES: Serum LH, FSH, E2, inhibin, P, and placental protein 14 (PP14) levels and histologic maturation of the endometrium. RESULTS: Serum FSH levels were increased whereas inhibin concentrations were reduced in the luteal-follicular transition of women > 40 years. No other hormonal changes were seen in this population, including P and PP14 secretion. Disruption of endometrial maturation occurred at a similar frequency in both age groups. CONCLUSIONS: Follicular recruitment, but not luteal function or endometrial maturation, is disturbed in cycling women > 40 years and may contribute to the decline in fertility with aging.
Assuntos
Envelhecimento/fisiologia , Endométrio/fisiologia , Glicoproteínas , Hormônios/sangue , Ciclo Menstrual/fisiologia , Adulto , Estradiol/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Glicodelina , Humanos , Inibinas/sangue , Hormônio Luteinizante/sangue , Pessoa de Meia-Idade , Proteínas da Gravidez/sangue , Progesterona/sangue , Estudos ProspectivosRESUMO
OBJECTIVE: To investigate whether a midluteal phase endometrial biopsy accurately predicts luteal function. DESIGN: One nonpregnant menstrual cycle was evaluated in a prospective fashion. SETTING: Outpatient Clinic of the Clinical Center of the National Institutes of Health. PARTICIPANTS: Fifty healthy, normally cycling women. INTERVENTIONS: Serum progesterone (P) was measured daily throughout the luteal phase. An endometrial biopsy was performed 7 to 9 days after the luteinizing hormone (LH) surge, as detected by rapid plasma assays, and dated histologically according to Noyes' criteria. MAIN OUTCOME MEASURE: To correlate endometrial maturation with luteal P secretion. RESULTS: Mean integrated P measurements were reduced only when the lag between histologic and chronological dating was > or = 3 days or > or = 4 days, depending on whether chronological dates were assigned prospectively from the LH surge or retrospectively from the onset of next menses, respectively. However, these lags did not consistently predict deficient luteal function because subnormal integrated P secretion was seen in only 14% of women with these delays in endometrial maturation. CONCLUSIONS: Midluteal phase endometrial biopsy provides a crude test of luteal function that does not precisely distinguish luteal insufficiency.
Assuntos
Corpo Lúteo/fisiologia , Endométrio/patologia , Fase Luteal , Adulto , Biópsia , Endométrio/crescimento & desenvolvimento , Feminino , Previsões , Humanos , Ciclo Menstrual , Pessoa de Meia-Idade , Progesterona/metabolismo , Fatores de TempoRESUMO
Mergers and health care agencies' response to mergers dominate current conversations in this evolving managed care environment. Hospitals are rapidly learning to adjust to declining occupancy rates and deceased utilization of resources. A business model to guide mergers was adapted to assist staff with the people, structural, cultural and political issues of organizational change. Creating successful new work environments, moving from a "We-they" mentality to unity and decreasing use of resources are outcomes described in this article.
Assuntos
Adaptação Psicológica , Instituições Associadas de Saúde/organização & administração , Reestruturação Hospitalar/organização & administração , Cultura Organizacional , Recursos Humanos em Hospital/psicologia , Humanos , Modelos Psicológicos , Inovação Organizacional , PolíticaRESUMO
1. During anaerobic glucose de-repression the respiration rate of whole cells of Saccharomyces carlsbergensis remained constant and was insensitive to antimycin A but was inhibited by 30% by KCN. Aeration of cells for 1 h led to increased respiration rate which was inhibited by 80% by antimycin A or KCN. 2. Homogenates were prepared from sphaeroplasts of anaerobically grown, glucose de-repressed cells and the distribution of marker enzymes was investigated after zonal centrifugation on sucrose gradients containing MgCl(2). These homogenates contained no detectable cytochrome c oxidase or catalase activity. The complex density distributions of NADH- and NADPH-cytochrome c oxidoreductases and adenosine triphosphatase(s) [ATPase(s)] were very different from those of anaerobically grown, glucose-repressed cells. 3. The specific activity of total ATPase was lowered and sensitivity to oligomycin decreased from 58 to 7% during de-repression. 4. Cytochrome c oxidase and catalase activities were detectable in homogenates of cells after 10min aeration. Zonal centrifugation indicated complex, broad sedimentable distributions of all enzyme activities assayed; the peaks of activity were at 1.27g/ml. 5. Centrifugation of homogenates of cells adapted for 30min and 3 h indicated a shift of density of the major sedimentable peak from 1.25g/ml (30min) to 1.235g/ml (3 h). After 30min adaptation a minor zone of oligomycin-sensitive ATPase and 15% of the total cytochrome c oxidase activities were detected at rho=1.12g/l; these particles together with those of higher density containing cytochrome c oxidase, ATPase and NADH-cytochrome c oxidoreductase activities were all sedimented at 10(5)g-min. 6. Electron microscopy indicated that the mitochondria-like structures of anaerobically grown, glucose-de-repressed cells were similar to those of repressed cells. After 10min of respiratory adaptation highly organized mitochondria were evident which resembled the condensed forms of mitochondria of aerobically grown, glucose-de-repressed cells. High-density zonal fractions of homogenates of cells after adaptation also contained numerous electron-dense vesicles 0.05-0.2mum in diameter. 7. The possibility that the ;promitochondria' of anaerobically grown cells may not be the direct structural precursors of fully functional mitochondria is discussed.
Assuntos
Glucose/metabolismo , Consumo de Oxigênio , Saccharomyces/enzimologia , Fosfatase Ácida/metabolismo , Adenosina Trifosfatases/metabolismo , Aerobiose , Anaerobiose , Antimicina A/farmacologia , Catalase/metabolismo , Centrifugação Zonal , Redutases do Citocromo/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Glucose/farmacologia , Microscopia Eletrônica , Saccharomyces/citologia , Saccharomyces/efeitos dos fármacos , Espectrofotometria , Espectrofotometria Ultravioleta , Fatores de TempoRESUMO
1. Homogenates were prepared from sphaeroplasts of aerobically grown glucose-de-repressed Saccharomyces carlsbergensis and the distributions of marker enzymes were investigated after differential centrifugation. Cytochrome c oxidase and cytochrome c were sedimented almost completely at 10(5)g-min, and this fraction also contained 37% of the catalase, 27% of the acid p-nitrophenyl phosphatase, 53 and 54% respectively of the NADH- and NADPH-cytochrome c oxidoreductases. 2. Zonal centrifugation indicated complex density distributions of the sedimentable portions of these enzymes and of adenosine triphosphatases and suggested the presence of two mitochondrial populations, as well as a bimodal distribution of peroxisomes and heterogeneity of the acid p-nitrophenyl phosphatase-containing particles. 3. Several different adenosine triphosphatases were distinguished in a post-mitochondrial supernatant that contained no mitochondrial fragments; these enzymes varied in their sensitivities to oligomycin and ouabain and their distributions were different from those of pyrophosphatase, adenosine phosphatase and adenosine pyrophosphatase. 4. The distribution of NADPH-cytochrome c oxidoreductase demonstrated that it cannot be used in S. carlsbergensis as a specific marker enzyme for the microsomal fraction. Glucose 6-phosphatase, inosine pyrophosphatase, cytochrome P-450 and five other enzymes frequently assigned to microsomal fractions of mammalian origin were not detected in yeast under these growth conditions.
Assuntos
Saccharomyces/citologia , Adenosina Trifosfatases/análise , Aerobiose , Catalase/análise , Fracionamento Celular , Centrifugação com Gradiente de Concentração , Centrifugação Zonal , Citocromos/análise , Complexo IV da Cadeia de Transporte de Elétrons/análise , Etanol/metabolismo , Glucose/metabolismo , Glucose-6-Fosfatase/análise , Inosina , Microscopia Eletrônica , NAD , NADP , Oxirredutases/análise , Monoéster Fosfórico Hidrolases/análise , Protoplastos , Pirofosfatases/análise , Saccharomyces/enzimologia , Saccharomyces/crescimento & desenvolvimento , Saccharomyces/metabolismo , Frações Subcelulares/enzimologiaRESUMO
1. Homogenates were prepared from sphaeroplasts of anaerobically grown, glucoserepressed Saccharomyces carlsbergensis, and the distributions of marker enzymes investigated after zonal centrifugation on sucrose gradients containing 2mm-MgCl(2). 2. These homogenates contained no detectable cytochrome c oxidase, succinate-cytochrome c oxidoreductase, succinate-ferricyanide oxidoreductase, l(+)-lactate-cytochrome c oxidoreductase or catalase. Cytochromes a+a(3) and c were not detected. 3. Zonal centrifugation of whole homogenates indicated complex density distributions of the sedimentable portions of NADH- and NADPH-cytochrome c oxidoreductases, adenosine triphosphatases (ATPases), adenosine pyrophosphatase (ADPase), pyrophosphatase and acid p-nitrophenyl phosphatase. Several different ATPases were distinguished on the basis of their sensitivities to oligomycin and ouabain. 4. Differential centrifugation of whole homogenates at 10(5)g-min left 80-90% of the protein, dithionite-reducible cytochrome b, acid hydrolases and pyrophosphatase in a supernatant (S(1)) together with 65 and 56% of the NADH- and NADPH-cytochrome c oxidoreductases respectively, 25% of the ATPases and 71% of the adenosine monophosphatase. 5. Further analysis of supernatant S(1) revealed the presence of a class of small particles containing NADPH-cytochrome c oxidoreductases and ATPases. 6. At least four different populations of large particles were distinguished. 7. Electron microscopy indicated that one of these corresponded to ;promitochondria' as described by other workers.
Assuntos
Fracionamento Celular , Saccharomyces , Adenosina Trifosfatases/análise , Anaerobiose , Centrifugação Zonal , Citocromos/análise , Glucose , Microscopia Eletrônica , NAD/análise , NADP/análise , Oxirredutases/análise , Frações Subcelulares/enzimologiaRESUMO
1. Subcellular fractionation of sphaeroplasts produced at different stages during the first 4h of respiratory adaptation of anaerobically grown glucose-de-repressed Saccharomyces carlsbergensis gave mitochondrial fractions that contained all the detectable c- and a-type cytochromes. 2. The rates of cytochrome formation were studied; individual cytochromes were produced at different rates so as to give respiratory chains having widely differing cytochrome ratios. A CO-reacting haemoprotein other than cytochrome a(3) also increased throughout 8h of respiratory adaptation. 3. Even after short periods of aeration, organisms contained mitochondria in which cytochrome-cytochrome interactions and the reaction of cytochrome a(3) with O(2) proceeded at rates almost as fast as in organelles from aerobically grown cells. 4. The technique of flow-flash photolysis enabled kinetic resolution of the reoxidation of cytochromes a(3) and a to be achieved and their individual contributions to extinction changes in the Soret region were assessed. The ratio cytochrome a(3)/cytochrome a increased over the early stages of adaptation.
Assuntos
Adaptação Fisiológica , Consumo de Oxigênio , Saccharomyces/metabolismo , Anaerobiose , Fracionamento Celular , Citocromos/metabolismo , Glucose/metabolismo , Cinética , Mitocôndrias/metabolismo , Organoides/metabolismo , Oxirredução , Fotólise , Fatores de TempoRESUMO
Adaptation of methanol-grown Candida boidinii to ethanol utilization was accompanied by an increase in proteolytic activities, which behaved like known vacuolar enzymes. Degradation of alcohol oxidase protein was partially prevented by the serine proteinase inhibitor phenylmethanesulphonyl fluoride, but not by the carboxyl proteinase inhibitor pepstatin. Fractionation of cell-free extracts, by high-speed zonal centrifugation, of methanol-grown C. boidinii showed non-sedimentable and sedimentable proteolytic activities. Naturally occurring inhibitors of vacuolar proteinases were non-sedimentable. Fractionation of extracts prepared from methanol-grown cells which had been adapted to ethanol utilization for 5 h revealed significant changes in the sedimentability and distribution of proteolytic and acid phosphatase activities. These results suggest the possible involvement of a vacuolar process during alcohol oxidase degradation.
Assuntos
Oxirredutases do Álcool/antagonistas & inibidores , Candida/enzimologia , Endopeptidases/metabolismo , Candida/efeitos dos fármacos , Centrifugação com Gradiente de Concentração , Cloroquina/farmacologia , Etanol/metabolismo , Metanol/metabolismo , Pepstatinas/farmacologia , Fluoreto de Fenilmetilsulfonil/farmacologia , Frações Subcelulares/enzimologiaRESUMO
A mixture of small (0.43-mum diameter) and large (0.62-mum diameter) low-density vesicles from spheroplasts of Saccharomyces cerevisiae was fractionated by rate centrifugation in a gradient of 0 to 8% (wt/vol) Ficoll to yield fractions rich (90 to 95%) in small or large vesicles. The large, but not small, vesicles swelled when diluted into mannitol solutions containing less than 0.4 M mannitol. The pH-electrophoretic mobility curve of the large vesicles showed that they are probably enclosed in a phospholipid-protein membrane. The dyes neutral red and toluidine blue, accumulated into large vesicles by intact cells and spheroplasts, were largely lost from large vesicles when these were separated from stained spheroplasts. Sudan black III stained small and large vesicles, both classes of vesicle retaining the stain on separation. Fractions rich in large vesicles contained proportionately more phospholipid and less free sterols, diacylglycerols, and free fatty acids compared with those enriched in small vesicles. The two classes of vesicles contained about the same proportions of esterified sterols and triacylglycerols. The free fatty acids in both small and large vesicles were free from unsaturated fatty-acyl residues; diacylglycerols and triacylglycerols contained appreciable proportions of unsaturated fatty-acyl residues. Small vesicles were richer in lipase activity, whereas the larger vesicles contained greater beta-glucanase and alpha-mannosidase activities. Phospholipase activity could not be detected in any of the fractions.
Assuntos
Saccharomyces cerevisiae/ultraestrutura , Fosfatase Ácida/metabolismo , Centrifugação com Gradiente de Concentração , Ácidos Graxos não Esterificados/análise , Glucana Endo-1,3-beta-D-Glucosidase/metabolismo , Glicerídeos/análise , Lipase/metabolismo , Lipídeos/análise , Manosidases/metabolismo , Organoides/análise , Organoides/enzimologia , Organoides/ultraestrutura , Fosfolipídeos/análise , Esferoplastos/ultraestrutura , Esteróis/análiseRESUMO
OBJECTIVE: Our objective was to determine if a progesterone antagonist might interdict the development of a secretory endometrium. STUDY DESIGN: Eleven normally cycling women not at risk for pregnancy received RU 486 (1 mg/day orally) or placebo throughout one menstrual cycle in a randomized, double-blind, crossover fashion. Estradiol, progesterone, and placental protein 14 were measured every 3 days; luteinizing hormone was measured until the midcycle surge was detected. An endometrial biopsy was performed on luteal phase day 7 to 9 and interpreted with Noyes' criteria. Differences between treatment groups were analyzed by the Student t test. RESULTS: RU 486 delayed ovulation, retarded endometrial maturation, and reduced peak levels of placental protein 14 without affecting gonadal steroid production. The abnormalities in endometrial morphology and function are similar to those seen in infertile women with luteal phase defects. CONCLUSION: We hypothesize that this regimen of antiprogestin administration may prevent implantation and offer a novel strategy for fertility control.
PIP: A small clinical study has provided preliminary evidence that a daily dose of 1 mg of RU-486 produces disruption of the morphology and function of the endometrium while preserving steroidogenesis, ovulation, and timing of the cycle. Enrolled in the study were 11 nonpregnant volunteers with regular menstrual cycles who received either RU-486 or a placebo during 2 treatment cycles in a randomized, crossover fashion. RU-486 delayed the midcycle luteinizing hormone surge and prolonged the follicular phase by 1-11 days in the 9 subjects included in the final analysis but did not alter the duration of the luteal phase. The mean length of the entire cycle increased an average of 6 days in RU-486 recipients. RU-486 also caused follicular phase estradiol and luteal phase progesterone concentrations to peak later compared with the placebo group. 6 of the 10 women who ovulated during RU-486 administration had delayed endometrial morphologic characteristics; another exhibited dyssynchrony between glandular and stromal tissue. Additional evidence for the antiprogestin effect of RU-486 on the endometrium was provided by finding of significantly lower peak placental protein 14 concentrations in treated subjects. RU-486 produced no effect on premenstrual symptoms, libido, dysmenorrhea, electrolytes, liver and renal functions, and cell blood counts. The potential of a small daily dose of a progesterone antagonist such as RU-486 as a fertility control agent merits further study.
Assuntos
Anticoncepcionais Orais , Endométrio/efeitos dos fármacos , Endométrio/fisiologia , Glicoproteínas , Mifepristona/farmacologia , Adulto , Método Duplo-Cego , Endométrio/anatomia & histologia , Estradiol/sangue , Feminino , Glicodelina , Humanos , Hormônio Luteinizante/sangue , Ovário/efeitos dos fármacos , Ovário/fisiologia , Ovulação/efeitos dos fármacos , Proteínas da Gravidez/sangue , Progesterona/sangue , Distribuição AleatóriaRESUMO
OBJECTIVE: Our purpose was to investigate the diagnostic accuracy of single or summed measurements of progesterone and placental protein 14, a progestin-dependent endometrial glycoprotein, in the evaluation of luteal function. STUDY DESIGN: Forty-five healthy women had daily blood measurements of luteinizing hormone, progesterone, and placental protein 14 during one menstrual cycle. RESULTS: Thirty-nine women had normal and six had deficient luteal function on the basis of serial progesterone determinations. Luteal insufficiency was not accurately diagnosed by single progesterone or placental protein 14 values or by integrated placental protein 14 measurements. In contrast, the condition was correctly identified in all but one cycle when the sum of progesterone on days 4 and 7 was < 49 nmol/L (15.4 ng/ml). A poor correlation was found between peak or integrated measurements of progesterone and placental protein 14. CONCLUSION: Measurement of serum progesterone, but not placental protein 14, on 2 days of the midluteal phase provides a convenient and reliable test of luteal function.